Lab 2 Aseptic Technique and Streak Plates...–Aseptic technique prevents contamination of sterile...
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1 Lab 2 – Aseptic Technique and Streak Plates The 5 I’s of Culturing Microbes • Inoculation • Incubation • Isolation • Inspection • Identification 1/10/2018 MDufilho
Transcript of Lab 2 Aseptic Technique and Streak Plates...–Aseptic technique prevents contamination of sterile...
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Lab 2 – Aseptic Technique and
Streak Plates The 5 I’s of Culturing Microbes
• Inoculation
• Incubation
• Isolation
• Inspection
• Identification
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Inoculation
Introduction of a small sample inoculum into a container of media to produce a culture of observable growth
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Culturing Microorganisms
• Obtaining Pure Cultures
– Cultures composed of cells arising from a single
progenitor
• Progenitor is termed a CFU
– Aseptic technique prevents contamination of
sterile substances or objects
– Two common isolation techniques
• Streak plates
• Pour plates
.
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Figure 6.9 Streak plate method of isolation-overview
Incubation
• Temperature-controlled chamber
• Microbe multiplies and produces
macroscopically observable growth
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Isolation
• If an individual bacterial cell is separated
from other cells and has space on a nutrient
surface, it will grow into a mound of cells--
a colony.
• A colony consists of one species.
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Figure 6.8 Characteristics of bacterial colonies-overview
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Inspection
Inspection – observation; macroscopic and microscopic
– Pure culture – grows only single known species of
microorganisms
– Mixed cultures – hold two or more identified species or
microorganisms
– Contaminated culture – once pure or mixed culture that
has unwanted microbes growing
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Various conditions of cultures
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Identification
Identification – macroscopic and microscopic
appearance, biochemical tests, genetic
characteristics, immunological testing
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Ex. 1.4 Aseptic Transfers
• Aseptic – What does it
mean?
• Inoculation Methods
• Inoculation Tools
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Sterilizing Instruments
Bunsen Burner Flame
Flaming Loop –
angle is wrong – should be
inverted
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Mixing Broth Cultures
Vortex Mixer Hand Mixing
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Aseptic Transfer
Removing Cap Flaming Tube
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Holding Tube
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Video
Broth to Broth Transfer
Move the tube/not loop Film on loop
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Plate inoculation Ex. 1.5
Invert Plate Streaking Plate
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Video
Streaking Plate
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